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Published on: March 24, 2020
Measurement of Postreplicative DNA Metabolism and Damage in the Rodent Brain
Jay P Patel1, Mark L Sowers1, Jason L Herring1
1Department of Pharmacology and Toxicology, ‡Department of Biochemistry and Molecular Biology, §Department of Anesthesiology, ∥Department of Internal Medicine, Division of Hematology and Oncology, ⊥Moody Project for Translational Traumatic Brain Injury Research, Department of Anesthesiology, University of Texas Medical Branch , Galveston, Texas 77555, United States.
Abstract:
The DNA of all organisms is metabolically active due to persistent endogenous DNA damage, repair, and enzyme-mediated base modification pathways important for epigenetic reprogramming and antibody diversity. The free bases released from DNA either spontaneously or by base excision repair pathways constitute DNA metabolites in living tissues. In this study, we have synthesized and characterized the stable-isotope standards for a series of pyrimidines derived from the normal DNA bases by oxidation and deamination. We have used these standards to measure free bases in small molecule extracts from rat brain. Free bases are observed in extracts, consistent with both endogenous DNA damage and 5-methylcytosine demethylation pathways. The most abundant free base observed is uracil, and the potential sources of uracil are discussed. The free bases measured in tissue extracts constitute the end product of DNA metabolism and could be used to reveal metabolic disturbances in human disease.
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